L-Carnitine拮抗培养心肌细胞缺氧/复氧损伤

目的 探讨L Carnitine对心肌细胞缺氧 /复氧损伤的防护作用及其可能的机制。方法 以原代培养新生鼠心肌细胞建立缺氧 /复氧损伤模型 ,观察不同剂量L Carnitine处理后细胞增殖活性、凋亡细胞百分率及DNA断片化率 ,作为反映L Carnitine对心肌细胞缺氧 /复氧损伤防护作用及其机制的指标。结果 在 5～ 5 0 0 0nmol/L范围内 ,L Carnitine预处理对培养心肌细胞缺氧 /复氧损伤均显示保护效应 ;在 5、5 0及 5 0 0nmol/L时均显著降低DNA断片化率 ;L Carnitine在 5 0nmol/L时与单纯缺氧 /复氧组比较可显著降低凋亡细胞百分率。结论 缺氧 /复氧对心肌细胞的影响之一是可诱导心肌细胞凋亡 ;L Carnitine对培养心肌细胞培养缺氧/复氧损伤具有防护作用 ,其机制可能与抑制缺氧 /复氧诱导细胞凋亡有关。

缺氧复氧对培养仔鼠心肌细胞NRF1和mtTFA表达的影响与L-carnitine保护作用

目的 探讨培养仔鼠心肌细胞缺氧复氧能量代谢损伤的机制及L 肉毒碱 (L carnitine )的保护作用。方法 以原代培养仔鼠心肌细胞建立缺氧复氧损伤模型 ,应用高效液相色谱检测心肌细胞内ATP、ADP、AMP含量 ;应用RT PCR方法检测心肌细胞核呼吸因子 (Nuclearrespiratoryfactor1 ,NRF1 )和线粒体转录因子A(MitochondrialtranscriptionfactorA ,mtTFA)mRNA表达。结果 与正常对照组比较 ,培养的心肌细胞缺氧 2 4h复氧 0、2、4、8h ,心肌细胞ATP、ADP含量均明显下降。与单纯缺氧复氧组比较 ,L carnitine预处理组心肌细胞缺氧复氧后ATP含量明显提升 ;培养的心肌细胞缺氧 2 4h ,NRF1和mtTFAmRNA明显降低 ,复氧 2、4、8h进一步降低。L carnitine预处理使缺氧 2 4h复氧 4h的心肌细胞NRF1和mtTFAmRNA表达分别提升 1 5 %和 2 0 %。结论 心肌细胞缺氧复氧过程中能量代谢明显降低 ,其损伤可能与NRF1和mtTFAmRNA表达下调有关 ;L carnitine对培养仔鼠心肌细胞缺氧复氧能量代谢损伤具有保护作用 ,其机制可能是L carnitine从转录调控水平改善了缺氧复氧过程中心肌细胞的能量代谢。

L-carnitine对缺氧/复氧新生大鼠心肌细胞能量生成及CPT-Ⅱ mRNA表达的影响

目的 探讨L 肉毒碱 (L carnitine ,L Car)对缺氧 复氧培养新生大鼠心肌细胞能量生成的影响及其机制。方法 以原代培养新生大鼠心肌细胞建立缺氧 复氧损伤模型 ,高效液相色谱检测单纯缺氧 复氧以及L Car预处理缺氧 复氧心肌细胞ATP、ADP、AMP生成改变 ;RT PCR方法检测心肌细胞肉毒碱脂酰基转移酶 Ⅱ (carnitinepalmitoyltransferase Ⅱ ,CPT Ⅱ )mRNA表达。结果 与正常对照组比较 ,培养心肌细胞缺氧 2 4h复氧 0、2、4、8h后 ,心肌细胞ATP、ADP均明显下降。与单纯缺氧 复氧组比较 ,L Car预处理组心肌细胞缺氧 复氧后ATP含量明显增高 ;培养心肌细胞缺氧 2 4h后 ,CPT ⅡmRNA表达除复氧 2h有所上升外 ,复氧 4、8h表达渐次降低。L Car预处理使缺氧 2 4h复氧 4h心肌细胞CPT ⅡmRNA表达呈浓度依赖性增高 ,5 0nmol L处理组CPT ⅡmRNA表达显著高于 0nmol L处理组。结论 心肌细胞缺氧 复氧过程中能量生成明显降低 ,其损伤可能与三羧酸循环中呼吸酶CPT Ⅱ表达降低有关 ;L Car对培养仔鼠心肌细胞缺氧复氧损伤具有显著保护作用 ,其机制可能是与L Car从转录调控水平改善CPT Ⅱ合成有关。

Beneficial effect of butyrate, Lactobacillus casei and L-carnitine combination in preference to each in experimental colitis

AIM: To investigate the beneficial effect of the combination of butyrate, Lactobacillus casei, and L-carnitine in a rat colitis model.

The Protective Effect of L-carnitine on Ischemia-reperfusion Heart

To investigate the protective effect of L-carnitine on myocardial ischemia-reperfusion injury in rat heart,all harvested isolated hearts were perfused on Langendorff apparatus with oxygenized K-H solution for 20 min. The hearts were then exposed to ischemia for 30 min. Following the ischemia the hearts were re-perfused with K-H solution for 120 min to serve as the control group A. Either 5 or 10 mmol/L of L-carnitine was added into the K-H solution for 20 min at the beginning of reperfusion to generate group B and group C, respectively. The derivatives of the intraventricular pressure curve （DP/DT）, left ventricular developed pressure （LVDP）, and coronary flux were monitored during the entire experiment. The levels of ATP, hepatin, malondialdehyde （MDA）, and superoxide dismutase （SOD） in tissue, and lactic dehydrogenase （LDH）, creatine phosphate kinase （CPK）, malondialdehyde （MDA）, and superoxide dismutase （SOD） concentration in the coronary efflux were all measured. Compared with the control group, the treatment with L-carnitine resulted in better results, i. e. , higher DP/DTmax and LVDP. At the same time, ventricular fibrillation was reduced, and the levels of ATP, hepatin and SOD were all elevated. However, the concentrations of MDA, CPK and LDH were all reduced. In conclusion, L-carnitine has a protective effect on ischemia-reperfusion injury, which is partly due to its prevention of energy loss and its antioxidant activity.

Enantiomeric Resolution on L-Carnitine Selective Polymers Prepared by Molecular Imprinting

L-carnitine selective polymers were prepared by molecular imprinting using methacrylic acid as the functional monomer. The acid function of the monomer is expected to form hydrogen bond and ionic interactions with the amine function of the target molecule L-carnitine. The imprinted polymers were used as stationary phases in high-performance liquid chromatography (HPLC). It was shown that L-carnitine imprinted polymer exhibited a higher affinity to its template molecule, while the non-imprinted polymer had no affinity to the compounds tested. Racemic carnitine hydrochloride was efficiently resolved on the L-carnitine imprinted polymer, and the separation factor is 1.9.

Development and validation of a UPLC–MS/MS assay for the determination of gemcitabine and its L-carnitine ester derivative in rat plasma and its application in oral pharmacokinetics

A simple and rapid UPLC–MS/MS method to simultaneously determine gemcitabine and its L-carnitine ester derivative(2’-deoxy-2’, 2’-difluoro-N-((4-amino-4-oxobutanoyl) oxy)-4-(trimethyl amm-onio) butanoate-cytidine, JDR) in rat plasma was developed and validated.The conventional plasma sample preparation method of nucleoside analogues is solidphase extraction(SPE) which is time-consuming and cost-expensive. In this study, gradient elution with small particles size solid phase was applied to effectively separate gemcitabine and JDR, and protein precipitation pretreatment was adopted to remove plasma protein and extract the analytes with high recovery(>81%). Method validation was performed as per the FDA guidelines, and the standard curves were found to be linear in the range of 5–4000 ng/ml for JDR and 4–4000 ng/ml for gemcitabine, respectively. The lower limit of quantitation(LLOQ)of gemcitabine and JDR was 4 and 5 ng/ml, respectively. The intra-day and inter-day precision and accuracy results were within the acceptable limits. Finally, the developed method was successfully applied to investigate the pharmacokinetic studies of JDR and gemcitabine after oral administration to rats.

Acetyl-L-carnitine:An Effective Antioxidant against Cryo-damage on Human Spermatozoa with Asthenospermia

A variety of natural and artificial cryoprotectant extenders have been explored to enhance sperm recovery following cryopreservation-thawing process. The current investigation is aimed at evaluating the effect of acetyl-L-carnitine on human spermatozoa and reactive species oxygen（ROS） level after freezing-thawing process. The spermatozoa were collected from 35 male patients diagnosed as having asthenospermia. The cryopreservation of human spermatozoa treated with acetyl-L-carnitine at different concentrations（group B： 2.5 mmol/L, group C： 7.5 mmol/L, group D： 15 mmol/L） was compared with control（group A： no acetyl-L-carnitine given）. For the frozen-thawed spermatozoa, the viability, motility and DNA integrity were measured by comet assay, acrosome integrity by FITC-PNA staining and ROS level was determined in each group. The results showed that there were no significant differences in motility and viability between group A and group B, while the motility and viability of spermatozoa in group C and group D were significantly increased as compared with those in group A. As compared with group A, the values for DNA integrity parameters including comet rate（CR）, tail DNA percentage（TD）, tail length（TL） and Oliver tail moment（OTM） were significantly reduced in group C and group D. Group C and group D also displayed a higher proportion of intact acrosome than group A. No significant difference in ROS level was found between group A and group B, while with the increase in acetyl-L-carnitine concentration, the ROS level in groups C and D was significantly reduced as compared with that in group A. In conclusion, acetyl-L-carnitine at a concentration of 7.5 mmol/L is an effective antioxidant against cryo-damage on post-thawed human spermatozoa.

L-carnitine alleviates sciatic nerve crush injury in rats:functional and electron microscopy assessments

Several studies have demonstrated that L-carnitine exhibits neuroprotective effects on injured sciatic nerve of rats with diabetes mellitus. It is hypothesized that L-carnitine exhibits neuro-protective effects on injured sciatic nerve of rats. Rat sciatic nerve was crush injured by a forceps and exhibited degenerative changes. After intragastric administration of 50 and 100 mg/kg L-carnitine for 30 days, axon area, myelin sheath area, axon diameter, myelin sheath diameter, and numerical density of the myelinated axons of injured sciatic nerve were similar to normal, and the function of injured sciatic nerve also improved signiifcantly. These ifndings suggest that L-carnitine exhibits neuroprotective effects on sciatic nerve crush injury in rats.

Effect of L-carnitine on Cardiomyocyte Apoptosis and Cardiac Function in Patients Undergoing Heart Valve Replacement Operation

Summary： The effects of L-carnitine, as an ingredient of cardioplegia solution, on cardiac function and cardiomyocyte apoptosis in patients undergoing heart valve replacement operation were investigated. Twenty-three cases undergoing heart valve replacement with cardiopulmonary bypass （CPB） were randomly allocated into two groups： L-carnitine group （n=12, 12 g/L L-carnitine was put in the ST. Thomas cardioplegia） and control group （n=11, identical to the L-carnitine group except that normal saline was administered instead of L-carnitine）. Serum cardial troponin I （cTnI） levels, the left ventricular ejection fraction （LVEF）, and cardiac index （CI） were measured perioperatively. A bit of myocardial tissue obtained from right atria was taken before CPB and by the end of intracardiac procedure to undergo electron microscopy examination and estimate apoptosis by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling （TUNEL）. From the end of CPB to 3 days after operation, the serum levels of cTnI in the L-carnitine group was significantly lower than that in the control group （P〈0.05）. Heart color ultrasonogram showed that the CI index and LVEF at 7th day postoperatively in the L-carnitine group were significantly higher than in the control group （P〈0.05）. Compared to the control group, L-carnitine significantly alleviated the morphologic changes of cardiac muscle cells （electron microscopy examination） and decreased the amounts of apoptotic cardiac muscle cells （TUNEL）. Furthermore, the dosage of vasoactive drugs used after operation was significantly less in the L-carnitine group （P〈0.01）. It was concluded that L-carnitine cardioplegia solution could improve cardiac function in patients undergoing heart valve replacement operation and alleviate CPB-mediated apoptosis of cardiac muscle cells.

L-carnitine supplementation in non-alcoholic fatty liver disease: A systematic review and meta-analysis

BACKGROUND Non-alcoholic fatty liver disease(NAFLD)dominates the landscape of modern hepatology.Affecting 25%of the general population,there is critical unmet need to identify broadly available,safe and cost-effective treatments.Cumulative evidence in animal and human models suggests that intrahepatic and skeletal muscle fatty acid oxidation is impaired in NAFLD,such that lipid accretion is not matched by efficient utilisation.L-carnitine is a crucial mediator of fatty acid metabolism in vivo,promoting mitochondrial lipidβ-oxidation and enhancing tissue metabolic flexibility.These physiological properties have generated research interest in L-carnitine as a potentially effective adjunctive therapy in NAFLD.AIM To systematically review randomised trials reporting effects of dietary L-carnitine supplementation on liver biochemistry,liver fat and insulin sensitivity in NAFLD.METHODS Search strategies,eligibility criteria and analytic methods were specified a priori(PROSPERO reference:CRD42018107063).Ovid MEDLINE,Ovid EMBASE,PubMed,Web of Science and the Cochrane Library were searched from their inception until April 2019.Outcome measures included serum concentrations of alanine and aspartate aminotransferase(ALT and AST),liver fat and insulin sensitivity assessed by the homeostasis model of insulin resistance(HOMA-IR).A random effects meta-analysis was performed for,ALT,AST and HOMA-IR measures separately.Between-study heterogeneity was measured using I2 statistics.RESULTS Five eligible randomised trials were included in the qualitative and quantitative synthesis(n=338).All of the 5 included trials assessed the effect of L-carnitine on serum ALT,identified from Italy,South Korea and Iran.Weighted mean difference(WMD)for ALT between L-carnitine and control groups after intervention was-25.34 IU/L[95%CI:-41.74-(-8.94);P=0.002].WMD for AST between L-carnitine and control groups was-13.68 IU/L(95%CI:-28.26-0.89;P=0.066).In three studies(n=204),HOMA-IR was evaluated.WMD for HOMA-IR between L-carnitine and control groups was-0.74 units[95%CI:-1.02-(-0.46);P<0.001].Two studies using validated outcome measures reported a significant reduction in liver fat in L-carnitine vs control groups post-intervention(P<0.001).CONCLUSION Pooled results indicate that L-carnitine supplementation attenuates ALT,liver fat and insulin resistance in NAFLD cohorts,confirming a beneficial effect of Lcarnitine for a highly prevalent condition with a growing economic burden.

Antioxidant Effects of L-carnitine on Rabbit

[ Objective] The paper was to study antioxidant effects of L-carnitine （LC） on rabbit, and explore its impact on SOD, GSH-PX, CAT, T-AOC and MDA activity in the plasma, brain tissue and cerebrospinal fluid of rabbit. [ Method] The healthy matured New Zealand rabbits with big ears were intragastrically administrated with LC at the dose of lmL/kg, and SOD, GSH-PX, CAT, T-AOC and MDA activities in plasma, brain tissue and cerebrospinal fluid of rabbit were determined at different periods before and after administration, respectively. [Result] After LC administration, SOD, GSH-PX CAT, and T-AOC activity in plasma, brain tissue and cerebrospinal fluid increased significantly （P 〈 0.05 ）, while MDA content decreased significantly （P 〈 0.05）. [Condusion ] LC oral solution shows antioxidant effects on rabbit.

Effects of L-carnitine on Blood Lipid Metabolism and Antioxidation in Hyperlipidemia Rats

[Objective] To study the effects of L-carnitine （LC） on lowering the high blood lipid and antioxidation in hyperlipidemia rats. [Method] 50 healthy SD rats were randomly divided into 5 groups. They were fed with standard diet, high-cholesterol diet and high-cholesterol diet with 0.25, 0.5, 1.0 g/（ kg · d） LC. After the LC groups were consecutively orally administered LC for 28 days, rat serum total cholesterol （ TC）, triglyceride （TG）, high density lipoprotein （ HDL-C）, low density lipoprotein （LDL-C） content, as well as super oxide dismutase （SOD） activity and malondial- dehyde （MDA） content in serum and liver were determined. [ Reset~ Compared with the high-fat model group, LC could significantly reduce the se- rum TC, TG, LDL-C levels, increase HDL-C level, and enhance SOD activity in serum and liver, decrease the content of MDA （ P 〈0.05）. [ Con- Clusion] LC might have a significant role in lowering the high blood lipid and improving internal antioxidant capacity in hyperlipidemia rats.

L-Carnitine Contents in the Tissues of Rabbits Fed Urea as an Alternative of Dietary Protein

The present study was aimed to observe the effects of urea ingestion, non-protein nitrogen, on the disorder of nitrogen metabolism with the L-carnitine contents using the blood, kidney, liver, and femoral muscle as markers. A total of 8 Japanese white rabbits were used in this experiment. They were fed a basal diet prepared for the control group and the nitrogen volume proportionated to one-third of CP 14%, was replaced with urea in the feed of the experimental group for 7 days. On the final day, the animals were fasted from the previous evening and sacrificed. Blood was collected into a test tube at the same time of the sacrifice and their heart, kidney, liver and femoral muscle were collected. The L-carnitine contents in each sample and the urea in the blood were determined. The results of the growth test showed that there was no significant difference. Furthermore, there was no significant difference in the contents of L-carnitine and urea in each sample. It was concluded that nitrogen replacement of the diet with urea, in the range of 1/3 of dietary protein, had neither effect on the maintenance of body weight nor nitrogen balance, including the de novo synthesis of L-carnitine.

The Possible Mechanisms Involved in the Protection Strategies against Radiation-Induced Cellular Damage by Carnitines

There is constant low level background radiation from the cosmos but in certain situation the body may be subjected to increased acute or chronic exposure from other sources. This occurs in situations such as radiation accidents, medical use and could possibly occur in military/terrorist incident. Dependent on the type, strength of the actual source, degree of exposure and type of radiation different strategies may be employed to reduce damage to the body tissues. A number of pharmacological agents such as peroxisome proliferator-activated receptor (PPAR) gamma agonists, diltiazem, amifostine and palifermin as well as antioxidants and metabolic compounds have been shown to be effective in preventing and also in reducing the long-term damage of the exposure of the living cells to radiation. The major drawback of synthetic (pharmacological) compounds has been that they are highly toxic at the optimum protective dose. Studies have shown that various endogenously found compounds such as L-carnitine, and its derivative acetyl-L-carnitine, are able to protect tissues and organs against various forms of toxic insult including radiation damage. The radiation-induced chronic injury may also be counteracted by other metabolic compounds with amine groups and antioxidant properties similar to the carnitines such as cysteine, 3,3’-diindolylmethane (DIM) and N-acetylcysteine. This review discuses the radioprotective compounds as well as the potential mechanism of cellular protection against radiation by carnitines and other compounds.

Effects of L-Carnitine on Propofol-Induced Inhibition of Free Fatty Acid Metabolism in Fasted Rats and in Vitro

Background: Propofol inhibits fatty acid oxidation and induces mitochondrial deficiency, a possible mechanism involved in propofol infusion syndrome. This study investigated how propofol influences fatty acid, glucose, and amino acid metabolism, as well as whether L-carnitine may improve suppression of free fatty acid metabolism. Methods: Male Sprague-Dawley rats, fasted for 16 hours, were allocated to the following two groups: (Group P;continuous intravenous administration of 10 mg/kg/h propofol;n = 8) and (Group P + C;intravenous administration of 50 mg/kg and then 50 mg/kg/h L-carnitine continuously;n = 8). Concentrations of glucose, free fatty acid (FFA), amino acids, in-sulin, and β-hydroxybutyric acid were measured at the start and then one, two, and three hours after propofol administration. Intrahepatic triglyceride levels were measured at the end of experiments. In vitro experiments comprised measurement of oxygen consumption in human hepatocytes (Hepg2) and investigating dependency on palmitic acid, glucose, and glutamine as fuel during propofol administration, with or without L-carnitine. Results: FFA increased in Group P and gradually decreased in Group P + C. There were significant differences between the two groups (Group P;331.2 ± 64.5 μM vs. Group P + C;199 ± 73.6 μM). Glucose decreased in both groups (Group P;53.8 ±16.6 mg/dL vs. Group P + C;88 ± 11.3 mg/dL). Amino acid concentrations were higher in Group P + C after experiments;alanine and glutamine increased significantly. β-hydroxybutyric acid increased significantly in Group P + C, and intrahepatic triglyceride decreased in Group P + C. Dependency on fatty acid metabolism significantly decreased with propofol only;addition of L-carnitine prevented these effects. Conclusions: Propofol impaired mitochondrial fatty acid metabolism, which was compensated mainly by a switch to glucose metabolism and partially by amino acid metabolism. Addition of L-carnitine may improve this imbalance of energy metabolism.

Behaviour and Degenerative Changes in the Basal Forebrain Systems of Aged Rats (12 Months Old) after Levo-Acetyl-Carnitine Treatments

One group of six male control rats [12 months old] and one group of six male rats of the same age, singularly maintained in a cage, and treated with acetyl-L-carnitine-HCl [(gamma-trimethyl-beta-acetyl-butyrobetaine-HCl: Sigma-Tau code ST200 or ALCAR: 60 mg/kg/day[7]/po)] for six months were tested in the spatial learning/memory Morris mazewater task and for atrophy and cell loss in seven myelo- and cytostructurally defined basal forebrain (BF) cholinergic regions [Freddi et al., 2009]. Coronal sections 25 ?m thick were cut through the BF regions and processed every 200 ?m for choline acetyltransferase (ChAT) immunohistochemistry. The ALCAR-treated rats had significantly shorter exit times on the Morris maze-water task test than the control rats (average ± SD 28.3 ± 12.4 s vs. 61.16 ± 4.67 s;t = 6.07, DOF = 10, P = 0.0001). Degenerative morphological changes in the BF ChAT-positive cells were observed in the substantia innominata pars anterior of the control rats but not in the treated animals (P < 0.05). In the BF, the counted and estimated average number of ChAT + cells in the 12-month-old ALCAR-treated rats (ChAT-ALCAR-12+ [Nos. 2,3,4]) was higher but not significantly (15.288 ± 3281) than that counted and estimated in the 12-month-old control rats [(ChAT-CT-12 [Nos. 1,2,3]) (11.508 ± 3868), t = 1.82, DOF = 10, P = 0.319]. In the substantia innominata pars posterior, the ChAT+ cells were significantly more numerous (P < 0.05) in the 12-month-old ALCAR-treated rats (ChAT-ALCAR-12 + [Nos. 2,3,4]) than in the control rats (ChAT-CT-12 [Nos. 1,2,3]). Above all, these results dem-onstrate that treatment with ALCAR from the age of 6 up to 12 months significantly attenuated spatial learning/memory impairment on the Morris maze-water behavioral task (P < 0.001) and also importantly reduced degeneration in size and number of cholinergic cells in the nucleus basalis magnocellularis of the BF. Accordingly, the surviving cholinergic neurons found in the BF of the ALCAR-treated rats might play an important role in modulating cortical activity and facilitating processes of attention, learning and memory.

Relationship between Serum L-Carnitine Levels and Sperm Parameters in Boars

This study evaluated the relationship between serum L-carnitine level and sperm parameters in young boars. Serum L-carnitine and semen characteristics were determined for 61 young Duroc boars between the ages of 590 and 630 days. Multiple linear regression analysis was performed to predict total and progressive motility and the total number of spermatozoa based on serum total L-carnitine and free L-carnitine levels. Total number of spermatozoa was not associated with basal serum L-carnitine levels. A regression equation was found in which both total L-carnitine levels and free L-carnitine levels were significant predictors of total and progressive motility (P 0.05). These results suggest that serum L-carnitine level is an important selection parameter for stock boars.

左旋肉毒碱(L-Carnitine)在能量代谢中的作用

详尽论述了左旋肉毒碱对猪与禽的能量代谢具有重要作用.在日粮中添加25～50 mg/kg左旋肉毒碱,可使仔猪日增重和饲料转化率提高,死亡率降低;产蛋鸡日粮中添加60 mg/kg左旋肉毒碱,可提高孵化率,并减少雏鸡死亡率;肉鸡日粮中添加50 mg/kg左旋肉毒碱,可促进肉鸡生长,降低瘁死率,改善鸡肉品质,提高饲料利用率.

非洲猪瘟病毒MGF360-13L蛋白的原核表达及多克隆抗体制备

本研究旨在通过构建非洲猪瘟病毒(ASFV)MGF360-13L基因原核表达系统表达13L蛋白,并制备其鼠源多克隆抗体。利用生物信息学方法,对ASFV MGF360-13L基因进行序列比对,分析其同源性、构建遗传进化树;将非洲猪瘟病毒MGF360-13L基因密码子优化后进行合成,连接至PET32a载体构建重组质粒pET32a-13L,转化至E.coliBL21(DE3)感受态细胞,经IPTG诱导表达获得目的蛋白,采用镍柱纯化法进行蛋白纯化。将纯化后的蛋白乳化后免疫8周龄BALB/c雌鼠制备多克隆抗体,间接ELISA和Western blot检测抗体特异性。SDS-PAGE结果显示,重组蛋白分子量大小为58.2 kDa,主要以包涵体形式存在;Western blot结果显示免疫猪阳性血清可特异性识别该蛋白,具有良好的反应性,表明该重组蛋白获得正确表达。利用该蛋白免疫小鼠制备多克隆抗体进行Western blot,结果显示其能与MGF360-13L重组蛋白发生特异性反应。间接ELISA测定抗体效价高达1∶256000。本研究成功制备非洲猪瘟病毒MGF360-13L重组蛋白,以其为免疫原制备的多克隆抗体具备较高的特异性和反应性,为进一步阐述MGF360-13L蛋白的生物学功能和研制非洲猪瘟新型疫苗奠定了基础。